Gong Ming, Ueda Yoshimichi, Kanazawa Yoshimitsu, Tsuchiya Hiroyuki, Ma Yi-gai
Department of Heamatology, China-Japan Friendship Hospital, Beijing 100029, China.
Zhonghua Zhong Liu Za Zhi. 2007 Jan;29(1):14-6.
To investigate the genes involved in pulmonary metastasis of human fibrosarcoma HT1080 cells in nude mice.
HT1080 cells were injected into the tail vein of BALB/ C nude mice. RNA samples were extracted from pulmonary metastatic tissues and normal control lung tissues, purified using Atlas Pure Total RNA labeling System (Clonetech Laboratories). cDNA probes labeled with 32P were prepared and hybridized to a cDNA membrane constructed with spots of 1176 human cancer related genes and radioactivities on the membrane were measeured by BAS 5000. The mRNA expression of gene FN1 was determined by real time RT-PCR using TaqMan methods. Furthermore, cells with FN1 expression were localized and obtained in situ in pulmonary metastatic foci by laser captured microdissection, and the FN1 expression was quantitated by real time RT-PCR.
Of the total 1176 genes, 27 genes (2. 3%) revealed to be apparently up-regulated and 4 genes (0. 3% ) down-regulated. Real time RT-PCR analysis verified significant up-regulation of gene FN1. Laser captured microdissection/ real time RT-PCR analysis demonstrated up-regulated gene FN1 not in stroma cells but in tumor cell nests.
Gene FN1 expression in fibrosarcoma HT1080 cells may be involved in pulmonary metastasis.
研究人纤维肉瘤HT1080细胞在裸鼠肺转移过程中涉及的基因。
将HT1080细胞注入BALB/C裸鼠尾静脉。从肺转移组织和正常对照肺组织中提取RNA样本，使用Atlas Pure Total RNA标记系统（Clonetech Laboratories公司）进行纯化。制备用32P标记的cDNA探针，并与由1176个人类癌症相关基因斑点构建的cDNA膜杂交，通过BAS 5000测量膜上的放射性。采用TaqMan方法通过实时RT-PCR测定基因FN1的mRNA表达。此外，通过激光捕获显微切割在肺转移灶中原位定位并获取具有FN1表达的细胞，并通过实时RT-PCR对FN1表达进行定量分析。
在总共1176个基因中，有27个基因（2.3%）明显上调，4个基因（0.3%）下调。实时RT-PCR分析证实基因FN1显著上调。激光捕获显微切割/实时RT-PCR分析表明，上调的基因FN1不在基质细胞中，而是在肿瘤细胞巢中。
纤维肉瘤HT1080细胞中的基因FN1表达可能参与肺转移。
